Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification

MR Bladergroen, KR Reiding, ALH Ederveen, GCM Vreeker, F Clerc, S Holst, Albert Bondt, M Wuhrer, YEM van der Burgt

Research output: Contribution to journalArticleAcademicpeer-review

69 Citations (Scopus)

Abstract

Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of alpha 2,6-linked sialic acid residues with parallel lactone formation of alpha 2,3-linked sialic acids. In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing.
Original languageUndefined/Unknown
Pages (from-to)4080-4086
Number of pages7
JournalJournal of Proteome Research
Volume14
Issue number9
DOIs
Publication statusPublished - 2015

Cite this